In Vitro Translation of mRNA in a Wheat Germ Extract Cell-Free System

  • Louise Olliver
  • Anne Grobler-Rabie
  • Charles D. Boyd
Part of the Springer Protocols Handbooks book series (SPH)


The wheat germ extract in vitro translation system has been used widely for faithful and efficient translation of viral and eukaryotic messenger RNAs in a heterologous cell-free system (1, 2, 3, 4, 5, 6, 7, 8, 9). With respect to the yield of translation products, the wheat germ extract is less efficient than most reticulocyte lysate cell-free systems (see  Chapters 106,  107, and  109). There are advantages, however, of using wheat germ extracts:
  1. 1.

    The in vivo competition of mRNAs for translation is more accurately represented, making the wheat germ system preferable for studying regulation of translation (1).

  2. 2.

    Particularly low levels of endogenous mRNA and the endogenous nuclease activity (10) obviate the requirement for treatment with a calcium-activated nuclease. There is, there fore, less disruption of the in vivo situation and contamination with calcium ions is less harmful. The identification of all sizes of exogenous mRNA-directed translation products is facilitated because of the low levels of endogenous mRNA present.

  3. 3.

    There is no posttranslational modification of translation products; primary products are therefore investigated, although processing may be achieved by the addition of microsomal membranes to the translation reaction.

  4. 4.

    The ionic conditions of the reaction may be altered to optimize the translation of large or small RNAs (2) (see Note 1).



Creatine Kinase Wheat Germ Translation Product Translation System Microsomal Membrane 
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  1. 1.
    Steward, A. G., Lloyd, M., and Arnstein, H. R. V. (1977) Maintenance of the ratio of α and β globin synthesis in rabbit reticulocytes. Eur. J. Biochem. 80, 453–459.CrossRefGoogle Scholar
  2. 2.
    Benveniste, K., Wilczek, J., Ruggieri, A., and Stern, R. (1976) Translation of collagen messenger RNA in a system derived from wheat germ. Biochemistry 15, 830–835.PubMedCrossRefGoogle Scholar
  3. 3.
    Huntner, A. R., Farrell, P. J., Jackson, R. J., and Hunt, T. (1977) The role of polyamines in cell-free protein in the wheat germ system. Eur. J. Biochem. 75, 149–157.CrossRefGoogle Scholar
  4. 4.
    Roberts, B. E. and Paterson, B. M. (1973) Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc. Natl. Acad. Sci. USA 70, 2330–2334.PubMedCrossRefGoogle Scholar
  5. 5.
    Davies, J. W., Aalbers, A. M. J., Stuik, E. J., and van Kammen, A. (1977) Translation of cowpea mosaic RNA in cell-free extract from wheat germ. FEBS Lett. 77, 265–269.PubMedCrossRefGoogle Scholar
  6. 6.
    Boedtker, H., Frischauf, A. M., and Lehrach, H. (1976) Isolation and translation of calva-ria procollagen messenger ribonucleic acids. Biochemistry 15, 4765–4770.PubMedCrossRefGoogle Scholar
  7. 7.
    Patrinou-Georgoulas, M. and John, H. A. (1977) The genes and mRNA coding for the theory chains of chick embryonic skeletal myosin. Cell 12, 491–499.PubMedCrossRefGoogle Scholar
  8. 8.
    Larkins, B. A., Jones, R. A., and Tsai, C. Y. (1976) Isolation and in vitro translation of zein messenger ribonucleic acid. Biochemistry 15, 5506–5511.PubMedCrossRefGoogle Scholar
  9. 9.
    Schroder, J., Betz, B., and Hahlbrock, K. (1976) Light-induced enzyme synthesis in cell suspension cultures of petroselinum. Eur. J. Biochem. 67, 527–541.PubMedCrossRefGoogle Scholar
  10. 10.
    Pelham, H. R. B. and Jackson, R. J. (1976) An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. J. Biochem. 67, 247–256.PubMedCrossRefGoogle Scholar
  11. 11.
    Darnbrough, C. H., Legon, S., Hunt, T., and Jackson, R. J. (1973) Initiation of protein synthesis: evidence for messenger RNA-independent binding of methionyl-transfer RNA to the 40S ribosomal subunit. J. Mol. Biol. 76, 379–403.PubMedCrossRefGoogle Scholar
  12. 12.
    Jackson, R. C. and Blobel, G. (1977) Post-translational cleavage of presecretory proteins with an extract of rough microsomes, from dog pancreas, with signal peptidase activity. Proc. Natl. Acad. Sci. USA 74, 5598–5602.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Louise Olliver
    • 1
  • Anne Grobler-Rabie
    • 1
  • Charles D. Boyd
    • 1
  1. 1.University of Medicine and DentistryNew Brunswick

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